Determination of pseudouridine in human urine and serum by high-performance liquid chromatography with post-column fluorescence derivatization.
A selective and sensitive method for the determination of pseudouridine in human urine and serum is described. The method is based on high-performance liquid chromatography with post-column fluorescence derivatization. Pseudouridine and 5-fluorouridine (internal standard) in a 10-fold diluted urine sample or a deproteinized serum sample are separated on a reversed-phase column (TSK gel ODS-80) with isocratic elution and successively subjected to derivatization involving periodate oxidation followed by fluorescence reaction with meso-1,2-bis(4-methoxyphenyl)ethylenediamine. The detection limit for pseudouridine is 4 pmol in a 100-microliters injection volume.
Umegae
Y
Y
Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Nohta
H
H
Ohkura
Y
Y
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005453:Fluorescence; D006801:Humans; D011560:Pseudouridine / Q000097:blood / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000037:antagonists & inhibitors; D014529:Uridine / Q000031:analogs & derivatives
D012335:RNA, Ribosomal; D011560:Pseudouridine; C001943:5-fluorouridine; D014529:Uridine